Molecular bases of cyclodextrin adapter interactions with engineered protein nanopores.

Journal article

Engineered protein pores have several potential applications in biotechnology: as sensor elements in stochastic detection and ultrarapid DNA sequencing, as nanoreactors to observe single-molecule chemistry, and in the construction of nano- and micro-devices. One important class of pores contains molecular adapters, which provide internal binding sites for small molecules. Mutants of the alpha-hemolysin (alphaHL) pore that bind the adapter beta-cyclodextrin (betaCD) approximately 10(4) times more tightly than the wild type have been obtained. We now use single-channel electrical recording, protein engineering including unnatural amino acid mutagenesis, and high-resolution x-ray crystallography to provide definitive structural information on these engineered protein nanopores in unparalleled detail.

Authors: Banerjee, A; Mikhailova, E; Cheley, S; Gu, L; Montoya, M

• Publication status: Published
• Journal: Proceedings of the National Academy of Sciences of the United States of America
• Volume: 107
• Issue: 18
• Pages: 8165-8170
• Publication date: 2010-05-01
• DOI: 10.1073/pnas.0914229107
• EISSN: 1091-6490
• ISSN: 0027-8424
• Language: English
• Keywords: Cyclodextrins; Recombinant Proteins; Crystallography, X-Ray; Protein Structure, Quaternary; Protein Structure, Tertiary; Hydrophobic and Hydrophilic Interactions; Thermodynamics; Models, Molecular; Porosity; Nanostructures; Kinetics; Mutation; Hemolysin Proteins